title: Rotavirus NSP1 mediates degradation of interferon regulatory factors through targeting of the dimerization domain.
abstract: Rotavirus nonstructural protein NSP1 can inhibit expression of interferon (IFN) and IFN-stimulated gene products by inducing proteasome-mediated degradation of IFN-regulatory factors (IRFs), including IRF3, IRF5, and IRF7. All IRF proteins share an N-terminal DNA-binding domain (DBD), and IRF3, IRF5, and IRF7 contain a similar C-proximal IRF association domain (IAD) that mediates IRF dimerization. An autoinhibitory domain (ID) at the extreme C terminus interacts with the IAD, burying residues necessary for IRF dimerization. Phosphorylation of serine/threonine residues in the ID induces charge repulsions that unmask the IAD, enabling IRF dimerization and subsequent nuclear translocation. To define the region of IRF proteins targeted for degradation by NSP1, we generated IRF3 and IRF7 truncation mutants and transiently expressed each with simian SA11-4F NSP1. These assays indicated that the IAD represented a necessary and sufficient target for degradation. Because NSP1 did not mediate degradation of truncated forms of the IAD, NSP1 likely requires a structurally intact IAD for recognition and targeting of IRF proteins. IRF9, which contains an IAD-like region that directs interactions with signal inducer and activator of transcription (STAT) proteins, was also targeted for degradation by NSP1, while IRF1, which lacks an IAD, was not. Analysis of mutant forms of IRF3 unable to undergo dimerization or that were constitutively dimeric showed that both were targeted for degradation by NSP1. These results indicate that SA11-4F NSP1 can induce degradation of inactive and activated forms of IAD-containing IRF proteins (IRF3 to IRF9), allowing a multipronged attack on IFN-based pathways that promote antiviral innate and adaptive immune responses.
authors.affiliation: Rotavirus Molecular Biology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Arnold
authors.forename: Mario
authors.initials: M
authors.lastname: Barro
authors.forename: John T
authors.initials: JT
authors.lastname: Patton
mesh_terms: D056704:Adaptive Immunity; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate; D050838:Interferon Regulatory Factor-3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D050839:Interferon Regulatory Factor-7 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D050835:Interferon Regulatory Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050834:Interferon-Stimulated Gene Factor 3, gamma Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012401:Rotavirus / Q000276:immunology* / Q000472:pathogenicity / Q000502:physiology; D017361:Viral Nonstructural Proteins / Q000276:immunology* / Q000378:metabolism*
chemical_list: C494232:IRF3 protein, human; C494234:IRF7 protein, human; C494227:IRF9 protein, human; D050838:Interferon Regulatory Factor-3; D050839:Interferon Regulatory Factor-7; D050835:Interferon Regulatory Factors; D050834:Interferon-Stimulated Gene Factor 3, gamma Subunit; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; C104282:nsp1 protein, Rotavirus